Epitope mapping of bovine retinal s-antigen with monoclonal antibodies

Volker Knospe, Larry A. Donoso, J. Paul Banga, Song Yue, Eva Kasp, Dale S Gregerson

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33 Scopus citations

Abstract

We examined the binding of seven murine monoclonal antibodies raised to S-antigen, an immunopathogenic, 404 residue photoreceptor cell autoantigen which induces experimental autoimmune uveoretinitis. S-antigen has also been identified as arrestin, a protein involved in the regulation of phototransduction. One additional monoclonal antibody (C10C10), raised to a synthetic peptide (peptide N) corresponding to residues 281 to 302 in bovine S-antigen, was also studied. In preliminary studies we examined the specificity of the antibody response to bovine S-antigen in sera from Balb/c mice. Western blots of mouse sera on the cyanogen bromide digest of bovine S-antigen demonstrated that all animals produced antibody which recognized epitopes within the C-terminal cyanogen bromide peptide designated CB46. Mice of the H-2d haplotype, including the Balb/c strain often used to produce monoclonal antibodies, showed little activity to cyanogen bromide peptides other than CB46. Also, all seven of the monoclonals raised to S-antigen are specific for epitopes in the CB46 peptide. The epitopes recognized by the monoclonal antibodies could be grouped into four distinct sites defined by peptides AE-1 (A2G5), peptide AA (PDS-1), peptide 19-OV (A9C6), and peptide 199 (BDS-1,2,3 and 4). The mono-clonal antibody, C10C1O, raised to peptide N recognizes an epitope in the N peptide and binds to a larger cyanogen bromide peptide designated CB123 as well as intact S-antigen. Fine mapping of these epitopes was done with various subpeptides. None of the antibodies bound the known immunopathogenic peptide, peptide M, which resides in CB123 although the C10C10 antibody binds a peptide adjacent to peptide M.

Original languageEnglish (US)
Pages (from-to)1137-1147
Number of pages11
JournalCurrent Eye Research
Volume7
Issue number11
DOIs
StatePublished - Jan 1 1988

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